Skip to main content
SpringerLink
Log in

Interaction of the cytoplasmic membrane and ribosomes in Escherichia coli; Altered ribosomal proteins in sucrose-dependent spectinomycin-resistant mutants

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Alterations in the ribosomes of sucrose-dependent spectinomycin-resistant (Sucd-Spcr) mutants of Escherichia coli were studied. Subunit exchange experiments showed that 30S subunits were responsible for the resistance of ribosomes to spectinomycin in all Sucd-Spcr mutants tested.

Proteins of 30S ribosomes were analyzed by carboxymethyl cellulose column chromatography based on their elution positions. Mutants YM22 and YM93 had an altered 30S ribosomal protein component, S5, and mutant YM50 had an altered protein, S4. Although a shift of elution position was not detected for all the 30S ribosomal proteins from mutant YM101, the amount of protein S3 was appreciably lowered in the isolated 30S subunits. A partial reconstitution experiment with protein S3 prepared from both the wild-type strain and YM101 revealed that the mutant had altered protein S3 which is responsible for the spectinomycin resistance.

These alterations in 30S subnits are discussed in relation to the interaction between ribosomes and the cytoplasmic membrane.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Bollen, A., Davies, J., Ozaki, M., Mizushima, S.: Ribosomal protein conferring sensitivity to the antibiotic spectinomycin in Escherichia coli. Science 165, 85–86 (1969)

    Google Scholar 

  • Dekio, S., Takata, R.: Genetic studies on the ribosomal proteins in Escherichia coli. II. Altered 30S ribosomal protein composition specific to spectinomycin resistant mutants. Molec. gen. Genet. 105, 219–224 (1969)

    Google Scholar 

  • Deusser, E., Stöffler, G., Wittmann, H.G.: Ribosomal proteins XVI. Altered S4 proteins in Escherichia coli revertants from streptomycin dependence to independence. Molec. gen. Genet. 109, 298–302 (1970)

    Google Scholar 

  • Funatsu, G., Schiltz, E., Wittmann, H.G.: Ribosomal proteins XXVII. Localization of the amino acid exchanges in protein S5 from two Escherichia coli mutants resistant to spectinomycin. Molec. gen. Genet. 114, 106–111 (1971)

    Google Scholar 

  • Held, W., Mizushima, S., Nomura, M.: Reconstitution of Escherichia coli 30S ribosomal subunits from purified molecular components. J. biolog Chem. 248, 5720–5730 (1973)

    Google Scholar 

  • Homann, H.E., Nierhaus, K.H.: Ribosomal proteins; protein compositions of biosynthetic precursors and artificial subparticles from ribosomal subunits in Escherichia coli K12. Europ. J. Biochem. 20, 249–257 (1971)

    Google Scholar 

  • Jaskunas, S.R., Nomura, M., Davies, J.: Genetics of bacterial ribosomes. Ribosomes (Nomura, M., Tissieres A., Lengyel, P. eds.), pp. 333–368 New York: Cold Spring Harbor Laboratory 1974

    Google Scholar 

  • Kreider, G., Brownstein B.L.: Ribosomal proteins involved in the suppression of streptomycin dependence in Escherichia coli. J. Bact. 109, 780–783, (1972)

    Google Scholar 

  • Lutter, L.C., Kurland, C.G., Stöffler, G.: Protein neighborhoods in the 30S ribosomal subunit of Escherichia coli. FEBS Letters 54, 144–150 (1975)

    Google Scholar 

  • Miyoshi, Y., Yamagata, H.: Sucrose-dependent spectinomycin-resistant mutants of Escherichia coli. J. Bact. 125, 142–148 (1976)

    Google Scholar 

  • Mizuno, T., Mizushima, S.: Characterization of stuck ribosomes induced by neomycin in vivo and in vitro. Biochim. biophys. Acta (Amst.) 353, 69–76 (1974)

    Google Scholar 

  • Mizuno, T., Yamada, H., Yamagata, H., Mizushima S.: Coordinated alterations in ribosomes and cytoplasmic membrane in sucrose-dependent, spectinomycin-resistant mutants of Escherichia coli. J. Bact. 125 524–530 (1976)

    Google Scholar 

  • Mizuno, T., Yamagata, H., Mizushima, S.: Interaction of cytoplasmic membrane and ribosomes in Escherichia coli: Spectinomycin induced disappearance of membrane protein I-19. J. Bact. 129, 326–332 (1977)

    Google Scholar 

  • Mizushima, S., Nomura, M.: Assembly mapping of 30S ribosomal proteins from E. coli. Nature (Lond.) 226, 1214–1218 (1970)

    Google Scholar 

  • Muto, A., Otaka, E., Itoh, T., Osawa, S., Wittmann, H.G.: Correlation of 30S ribosomal proteins of Escherichia coli fractionated on carboxymethyl-cellulose column chromatography to the standard nomenclature. Molec. gen. Genet. 140, 1–5 (1975)

    Google Scholar 

  • Nashimoto, H., Uchida, H.: Late steps in the assembly of 30S ribosomal proteins in vivo in a spectinomycin-resistant mutant of Escherichia coli. J. molec. Biol. 96, 443–453 (1975)

    Google Scholar 

  • Otaka, E., Itoh, T., Osawa, S.: Ribosomal proteins of bacterial cells; strain- and species-specificity. J. molec. Biol. 33, 93–107 (1968)

    Google Scholar 

  • Ozaki, M., Mizushima, S., Nomura, M.: Identification and functional characterization of the protein controlled by the streptomycin-resistant locus in E. coli. Nature (Lond.) 222, 333–339 (1969)

    Google Scholar 

  • Peretz, H., Towbin, H., Elson, D.: The use of a cleavable crosslinking reagent to identify neighboring proteins in the 30S ribosomal subunit of Escherichia coli Europ. J. Biochem. 63, 83–92, (1976)

    Google Scholar 

  • Sabatini, D.D., Tashiro, Y., Palade, G.E.: On the attachment of ribosomes to microsomal membranes. J. molec. Biol. 19, 503–524 (1966)

    Google Scholar 

  • Sommer, A., Traut, R.R.: Identidication of neighboring protein pairs in the Escherichia coli 30S ribosomal subunit by crosslinking with methyl-4-mercaptobutyrimidate. J. molec. Biol. 106, 995–1015 (1976)

    Google Scholar 

  • Tischendorf, G.W., Zeichhardt, H., Stöffler, G.: Architecture of the Escherichia coli ribosome as determined by immune electron microscopy. Proc. nat. Acad. Sci. (Wash.) 72, 4820–4824 (1975)

    Google Scholar 

  • Traub, P., Mizushima S., Lowry, C.V., Nomura, M.: Reconstitution of ribosomes from subribosomal components. In: Methods in enzymol. (Moldave, K., Grossman, L. eds.), Vol. 20, pp. 391–407. New York: Academic Press 1971

    Google Scholar 

  • Traub, P., Nomura, M.: Structure and function of Escherichia coli ribosomes. VI. Mechanism of assembly of 30S ribosomes studied in vitro. J. molec. Biol. 40, 391–413 (1969)

    Google Scholar 

  • Wallace, B.J., Phang-Cheng, T., Davis, B.D.: Selective inhibition of initiating ribosomes by spectinomycin Proc. nat. Acad. Sci. (Wash.) 71, 1634–1638 (1974)

    Google Scholar 

Download references

Author information

Author notes

  1. Takeshi Mizuno

    Present address: Mitsubishi-Kasei Institute of Life Science, 194, Machida, Tokyo, Japan

Authors and Affiliations

  1. Department of Agricultural Chemistry, Nagoya University, Chikusa-ku, 464, Nagoya, Japan

    Munehiko Dombou, Takeshi Mizuno & Shoji Mizushima

Authors
  1. Munehiko Dombou
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Takeshi Mizuno
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shoji Mizushima
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by H. G. Wittmann

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dombou, M., Mizuno, T. & Mizushima, S. Interaction of the cytoplasmic membrane and ribosomes in Escherichia coli; Altered ribosomal proteins in sucrose-dependent spectinomycin-resistant mutants. Molec. Gen. Genet. 155, 53–60 (1977). https://doi.org/10.1007/BF00268560

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00268560

Keywords

Search

Navigation